We propose to use the honey bee as a model to determine which of the mechanisms that can be manipulated to increase lifespan actually have been in nature. The queen bee is highly reproductively active but typically lives 10-fold longer than does the worker bee. Using the free radical damage theory of aging as a foundation, we will: 1) Determine whether queen-worker longevity differences are associated with differences in expression of genes encoding antioxidants, electron chain proteins, or both. We have complete or near complete sequence for 25 genes related to longevity: 15 antioxidant, 8 mitochondrial, and 2 signal transduction genes; 12 will be analyzed with real-time quantitative RT-PCR and the rest are on a Cdna microarray (below). Analyses will be supplemented with protein measures in selected cases. Preliminary results indicate striking differences in gene expression, especially early in life. Measurements of ATP production and effects of ROS damage will test the functional significance of the differences; 2) Determine the causal basis of some of the queen-worker differences in gene expression with transgenic flies. Three of the genes with some of the strongest queen-worker differences will be selected and transgenic flies made (up- and downregulation) in the laboratory of collaborator J. Tower. Collaborator K. Hughes will study their age-specific survival and reproduction, and developmental rate. Hughes will also work with long- and-short lived selected lines of flies to determine whether selection acted on some of the same genes that differ in queens vs. workers; and 3) Conduct a microarray survey to identify additional genes that differ in expression between queens and workers, to determine what related pathways are affected in association with differences in the specific genes studied in Aim 1. We will use our recently developed cDNA microarrays which represent ca. 6000 different bee genes, including additional antioxidant and respiration-related genes, and genes encoding different HSPs. The principal significance of this research is that it will identify naturally occurring molecular mechanisms promoting longevity. The value of the bee as an aging model will be enhanced significantly with the expected completion of genome sequencing later in '03.